Assignment2 - Differential Gene expression and Preliminary ORA
Zhiwen Tan, 1003905287
- Assignment 1 summary
- Differential Gene Expression
- Thresholded over-representation analysis
- Threshold list
- Which method did you choose and why?
- What annotation data did you use and why? What version of the annotation are you using?
- How many genesets were returned with what thresholds?
- Run the analysis using the up-regulated set of genes, and the down-regulated set of genes separately. How do these results compare to using the whole list (i.e all differentially expressed genes together vs. the up-regulated and down regulated differentially expressed genes separately)?
- Threshold list
- Interpretation
- Reference
Assignment 1 summary
install all packages if needed, then library them. if file normalized_data.rds and filtered_data.rds does not exist, rerun A1.
#install Biocmanager in order to install necessary package
if (!requireNamespace("BiocManager", quietly = TRUE))
install.packages("BiocManager")
#install necessary package if not installed
if (!requireNamespace("GEOmetadb", quietly = TRUE))
BiocManager::install("GEOmetadb")
if (! requireNamespace("GEOquery", quietly = TRUE)) {
BiocManager::install("GEOquery")
}
if (! requireNamespace("edgeR", quietly = TRUE)) {
BiocManager::install("edgeR")
}
# install limma if not exist
if (! requireNamespace("limma", quietly = TRUE)) {
BiocManager::install("limma")
}
if (! requireNamespace("ComplexHeatmap", quietly = TRUE)) {
BiocManager::install("ComplexHeatmap")
}
if (! requireNamespace("circlize", quietly = TRUE)) {
install.packages("circlize")
}
library(dplyr)
library(GEOmetadb)
library(GEOquery)
library(edgeR)
library(knitr)
library(RSQLite)
library(limma)
library(ComplexHeatmap)
library(circlize)
# load all data from A1
if (! file.exists("./normalized_data.rds") || ! file.exists("filtered_data.rds")){
rmarkdown::render("BCB420_A1.Rmd")
}First, we want to get the description for dataset .
# First, we want to get the description for dataset GSE145412.
gse <- getGEO("GSE145412",GSEMatrix=FALSE)
kable(data.frame(head(Meta(gse))), format = "html")Then we want to show more information associated with the dataset.
# Then, we want to get information about the dataset
current_gpl <- names(GPLList(gse))[1]
current_gpl_info <- Meta(getGEO(current_gpl))## File stored at:## /tmp/Rtmpi6Z4MH/GPL18573.softThe first assignment was focused on clean up data, grouping, and using TMM method to normalized the dataset. the results are shown in both boxplot and density plot.
normalized_data = readRDS("normalized_data.rds")
# boxplot for normalized data
data2plot_after <- log2(normalized_data)
{boxplot(data2plot_after, xlab = "Samples", ylab = "log2 CPM",
las = 2, cex = 0.5, cex.lab = 0.5,
cex.axis = 0.5, main = "MetS/obese RNASeq Samples normalized")
abline(h = median(apply(data2plot_after, 2, median)), col = "green", lwd = 0.6, lty = "dashed")}
# density plot for normalized data
norm_counts_density <- apply(log2(normalized_data), 2, density)
xlim <- 0; ylim <- 0
for (i in 1:length(norm_counts_density)) {
xlim <- range(c(xlim, norm_counts_density[[i]]$x));
ylim <- range(c(ylim, norm_counts_density[[i]]$y))
}
cols <- rainbow(length(norm_counts_density))
ltys <- rep(1, length(norm_counts_density))
# plot the second density plot
plot(norm_counts_density[[1]], xlim=xlim, ylim=ylim, type="n",
ylab="Smoothing density of log2-CPM", main="MetS/obese RNASeq Samples normalized",
cex.lab = 0.85)
# plot each line
for (i in 1:length(norm_counts_density))
lines(norm_counts_density[[i]], col=cols[i], lty=ltys[i])
In this assignment, those genes will be ranked according to differential expression and I will perform enrichment analysis using g:Profiler to see the dominant themes in the top hits. From the results, I will determine if my study supports the original finding.
Differential Gene Expression
Loading normalized data
get the normalized data, and convert it to numerical matrix
# take a look at the data
kable(normalized_data[1:5,1:5], type="html")| sample_1 | sample_4 | sample_5 | sample_6 | sample_7 | |
|---|---|---|---|---|---|
| 7SK | 1.8665112 | 0.4927562 | 2.3163723 | 0.6257193 | 0.4934407 |
| A1BG-AS1 | 0.2201429 | 0.2269133 | 0.2236347 | 0.1269422 | 0.1814833 |
| A2M-AS1 | 0.0419011 | 0.0575317 | 0.0302224 | 0.0658098 | 0.0579427 |
| A3GALT2 | 0.0448311 | 0.0154599 | 0.0187952 | 0.0399286 | 0.0213537 |
| AAAS | 0.4379074 | 0.4786196 | 0.4105074 | 0.5047566 | 0.4477859 |
heatmap_matrix <- as.matrix(normalized_data)Heatmap and Model
install required package and library them, then show the heat map
Show the heat map
current_heatmap
Use limma to draw the MDSplot
# MDS plot for the heat map
plotMDS(heatmap_matrix, col = rep(c("darkgreen","blue"),10))
# Draw MDS plot color by patient to see how samples are clustering
pat_colors <- rainbow(4)
pat_colors <- unlist(lapply(pat_colors,FUN=function(x){rep(x,2)}))
plotMDS(heatmap_matrix, col = pat_colors)
Now we want to introduce groups
# Now define groups
samples <- data.frame("type" = c("O", "L", "L", "O", "O", "L", "O",
"L", "L", "O", "L", "L", "L", "L",
"O", "O", "L", "O", "O", "O", "L",
"O", "L", "O", "L", "O", "L", "O",
"O", "L", "L", "O"),
"disease" = c("H", "H", "D", "D", "D", "D", "H",
"H", "H", "D", "H", "H", "D", "D",
"D", "D", "D", "H", "H", "H", "H",
"H", "D", "D", "H", "H", "H", "H",
"D", "D", "D", "D"),
"patients" = c("sample_1", "sample_4", "sample_5",
"sample_6", "sample_7", "sample_8",
"sample_9", "sample_10", "sample_11",
"sample_13", "sample_15", "sample_17",
"sample_18", "sample_19", "sample_21",
"sample_22", "sample_24", "sample_25",
"sample_28", "sample_29", "sample_32",
"sample_33", "sample_34", "sample_35",
"sample_36", "sample_39", "sample_42",
"sample_43", "sample_44", "sample_46",
"sample_47", "sample_48"))
# Add corresponding rownames to each cell
rownames(samples) <- colnames(normalized_data)[1:32]
samples <- data.frame(samples)
samples$name <- paste(samples$disease, samples$type, samples$patients)
colnames(heatmap_matrix) <- samples$name
colnames(normalized_data) <- samples$name
# Show the first 5 rows
samples[1:5,]Create model matrix, and fit normalized data into the model, only used disease (H - healthy, D - disease) column for analysis
# set model to only disease
model_design <- model.matrix(~ samples$disease)
kable(model_design[1:5,], type="html")| (Intercept) | samples$diseaseH |
|---|---|
| 1 | 1 |
| 1 | 1 |
| 1 | 0 |
| 1 | 0 |
| 1 | 0 |
expressionMatrix <- as.matrix(normalized_data)
minimalSet <- ExpressionSet(assayData=expressionMatrix)
fit <- lmFit(minimalSet, model_design)Apply empirical Bayes to compute differential expression
fit2 <- eBayes(fit,trend=TRUE)
output_hits <- topTable(fit2, coef=ncol(model_design), adjust.method = "BH",
number = nrow(expressionMatrix))
kable(output_hits[1:5,],type="html")| logFC | AveExpr | t | P.Value | adj.P.Val | B | |
|---|---|---|---|---|---|---|
| RP11-517B11.7 | 0.0118532 | 0.0339949 | 4.743916 | 0.0000435 | 0.4633653 | 1.8276821 |
| CACNA1C-AS1 | 0.0249612 | 0.0814327 | 4.279405 | 0.0001637 | 0.4633653 | 0.7380255 |
| CDKN2D | -1.5100146 | 5.5504830 | -4.233263 | 0.0001865 | 0.4633653 | 0.6304486 |
| ARPC1A | -0.2093313 | 1.4309654 | -4.065386 | 0.0002991 | 0.4633653 | 0.2407121 |
| ALOX12 | -0.1625949 | 0.2339444 | -3.938615 | 0.0004261 | 0.4633653 | -0.0515034 |
check how many genes has p value less than 0.05, and how many genes pass the correction
length(which(output_hits$P.Value < 0.05))## [1] 1630length(which(output_hits$adj.P.Val < 0.05))## [1] 0Question 1:
There are 1630 genes were significantly deferentially expressed, the thresholds I used was 0.05 because it is the most widely accepted p-value, and it means there is a less than 5% chance the result could be random.
there is no gene passes the correction, so now we will try to improve the result.
Multiple Hypothesis Correction
now we need to correct the p-value for my data using multiple hypothesis correction.
# creates a design matrix with the consideration of both body type and disease
model_design_pat <- model.matrix( ~ samples$type + samples$disease)
kable(model_design_pat,type="html")| (Intercept) | samples\(typeO| samples\)diseaseH | |
|---|---|---|
| 1 | 1 | 1 |
| 1 | 0 | 1 |
| 1 | 0 | 0 |
| 1 | 1 | 0 |
| 1 | 1 | 0 |
| 1 | 0 | 0 |
| 1 | 1 | 1 |
| 1 | 0 | 1 |
| 1 | 0 | 1 |
| 1 | 1 | 0 |
| 1 | 0 | 1 |
| 1 | 0 | 1 |
| 1 | 0 | 0 |
| 1 | 0 | 0 |
| 1 | 1 | 0 |
| 1 | 1 | 0 |
| 1 | 0 | 0 |
| 1 | 1 | 1 |
| 1 | 1 | 1 |
| 1 | 1 | 1 |
| 1 | 0 | 1 |
| 1 | 1 | 1 |
| 1 | 0 | 0 |
| 1 | 1 | 0 |
| 1 | 0 | 1 |
| 1 | 1 | 1 |
| 1 | 0 | 1 |
| 1 | 1 | 1 |
| 1 | 1 | 0 |
| 1 | 0 | 0 |
| 1 | 0 | 0 |
| 1 | 1 | 0 |
# fit our data
fit_pat <- lmFit(minimalSet, model_design_pat)
# Apply empircal Bayes to compute differential expression
fit2_pat <- eBayes(fit_pat,trend=TRUE)
topfit_pat <- topTable(fit2_pat, coef=ncol(model_design_pat),
adjust.method = "BH",
number = nrow(expressionMatrix))
output_hits_pat <- topfit_pat[order(topfit_pat$P.Value),]
kable(output_hits_pat[1:5,], type="html")| logFC | AveExpr | t | P.Value | adj.P.Val | B | |
|---|---|---|---|---|---|---|
| RP11-517B11.7 | 0.0118532 | 0.0339949 | 4.938247 | 0.0000269 | 0.4183861 | 2.2350673 |
| ARPC1A | -0.2093313 | 1.4309654 | -4.359947 | 0.0001377 | 0.4378044 | 0.8915791 |
| CACNA1C-AS1 | 0.0249612 | 0.0814327 | 4.221639 | 0.0002027 | 0.4378044 | 0.5719516 |
| CDKN2D | -1.5100146 | 5.5504830 | -4.171077 | 0.0002334 | 0.4378044 | 0.4554521 |
| ZNF880 | 0.0238263 | 0.1217908 | 4.002758 | 0.0003722 | 0.4378044 | 0.0693330 |
check how many genes has p value less than 0.05, and how many genes pass the correction
length(which(output_hits_pat$P.Value < 0.05))## [1] 1715length(which(output_hits_pat$adj.P.Val < 0.05))## [1] 0Question 2:
I choose to use the BH (Benjamini-Hochberg) method because it is a popular and powerful method for controlling the false discovery rate in the field of science. The number of genes that has p value less than 0.05 increased to 1715, but the number of genes pass correction is still 0.
now, we want to compare the two results by plot
# define colors
simple_model_pvalues <- data.frame(id = rownames(output_hits),
simple_pvalue=output_hits$P.Value)
pat_model_pvalues <- data.frame(id = rownames(output_hits_pat),
patient_pvalue = output_hits_pat$P.Value)
two_models_pvalues <- merge(simple_model_pvalues,
pat_model_pvalues,by.x=1,by.y=1)
two_models_pvalues$colour <- "black"
two_models_pvalues$colour[two_models_pvalues$simple_pvalue<0.05] <- "orange"
two_models_pvalues$colour[two_models_pvalues$patient_pvalue<0.05] <- "blue"
two_models_pvalues$colour[two_models_pvalues$simple_pvalue<0.05 &
two_models_pvalues$patient_pvalue<0.05] <- "red"
# Plotting
plot(two_models_pvalues$simple_pvalue, two_models_pvalues$patient_pvalue,
col = two_models_pvalues$colour, xlab = "simple model p-values",
ylab ="Patient model p-values", main="Simple vs Patient Limma")
# add legend
legend(0, 1,legend=c("simple", "patient", "both", "other"),
col=c("orange", "blue", "red", "black"), lty=1:2, cex=0.9)
The plot looks like a straight line, this suggest that the differential expression for both simple model and patient model gives similar result. the body type of the patients does not effect the result much.
The above result was come from limma, now we want to know the result using edgeR
filtered_data = readRDS("filtered_data.rds")
rownames(filtered_data) <- filtered_data$Group.1
# create a EdgeR object
d = DGEList(counts=as.matrix(filtered_data[2:33]), group=samples$disease)
# create model
model_design_pat <- model.matrix( ~ samples$type + samples$disease)
# estimate dispersion
d <- estimateDisp(d, model_design_pat)
# calculate normalization factors
d <- calcNormFactors(d)
# fit model
fit <- glmQLFit(d, model_design_pat)
# calculate differential expression
qlf.hea_vs_dis <- glmQLFTest(fit)
# get all results
qlf_output_hits <- topTags(qlf.hea_vs_dis,sort.by = "PValue",
n = nrow(filtered_data))
length(which(qlf_output_hits$table$PValue < 0.05)) # 136## [1] 136length(which(qlf_output_hits$table$FDR < 0.05)) # 0 ## [1] 0kable(topTags(qlf.hea_vs_dis), type="html")
|
|
|
|
# check how many genes are up regulated
length(which(qlf_output_hits$table$PValue < 0.05 &
qlf_output_hits$table$logFC > 0)) # 5## [1] 5# check how many genes are down regulated
length(which(qlf_output_hits$table$PValue < 0.05 &
qlf_output_hits$table$logFC < 0)) # 131## [1] 131This means no gene has a FDR value less than 0.05, and only 5 genes are up regulated and 131 genes are down regulated
Compare limma with Quasi liklihood
qlf_pat_model_pvalues <- data.frame(
ensembl_id = rownames(qlf_output_hits$table),
qlf_patient_pvalue=qlf_output_hits$table$PValue)
limma_pat_model_pvalues <- data.frame(
ensembl_id = rownames(output_hits_pat),
limma_patient_pvalue = output_hits_pat$P.Value)
two_models_pvalues <- merge(qlf_pat_model_pvalues,
limma_pat_model_pvalues,
by.x=1,by.y=1)
two_models_pvalues$colour <- "black"
two_models_pvalues$colour[two_models_pvalues$qlf_patient_pvalue<0.05] <- "orange"
two_models_pvalues$colour[two_models_pvalues$limma_patient_pvalue<0.05] <- "blue"
two_models_pvalues$colour[two_models_pvalues$qlf_patient_pvalue<0.05 & two_models_pvalues$limma_patient_pvalue<0.05] <- "red"plot(two_models_pvalues$qlf_patient_pvalue,
two_models_pvalues$limma_patient_pvalue,
col = two_models_pvalues$colour,
xlab = "QLF patient model p-values",
ylab ="Limma Patient model p-values",
main="QLF vs Limma")
legend(0, 0.9,legend=c("QLF", "Limma", "both QLF and Limma", "other"),
col=c("orange", "blue", "red", "black"), lty=1:2, cex=0.6)
MA plot
Show the amount of deferentially expressed genes using an MA Plot. I split the data into two groups, one is Healthy group and one is disease group. Since plotMD need to column of data, I calculated the average for each gene in both groups. Then I need to make sure using the right color for significant data, so I reordered the topfit_pat and output_hits. Then I need to define what gene to color, so I introduced a status for coloring. After all the process, I can make the MA plot for my data.
group1 <- normalized_data[, grepl("H O", names(normalized_data))]
group1 <- data.frame(Means=rowMeans(group1))
group2 <- normalized_data[, grepl("D O", names(normalized_data))]
group2 <- data.frame(Means=rowMeans(group2))
combined_group <- cbind(group1, group2)
temp_hit_pat <- topfit_pat[order(rownames(topfit_pat)),]
temp_hit <- output_hits[order(rownames(output_hits)),]
status <- rep(0, nrow(normalized_data))
status[temp_hit$logFC > 0 & temp_hit_pat$P.Value < 0.05] <- 1
status[temp_hit$logFC < 0 & temp_hit_pat$P.Value < 0.05] <- -1
plotMD(log2(combined_group[, c(1, 2)]), status=status, values=c(1,-1), hl.col=c("blue", "red"), ylab="M - ratio log expression", main="Healthy vs Disease", )
Question3:
The gene of interest are highlighted in red and blue colors in the MA plot.
Visualize tophits using heatmap
redraw the heat map with the condition p.value < 0.05
top_hits <- rownames(output_hits_pat)[output_hits_pat$P.Value < 0.05]
heatmap_matrix_tophits <- scale(heatmap_matrix[which(rownames(heatmap_matrix) %in% top_hits),])
if(min(heatmap_matrix_tophits) == 0){
heatmap_col = colorRamp2(c( 0, max(heatmap_matrix_tophits)),
c( "white", "red"))
} else {
heatmap_col = colorRamp2(c(min(heatmap_matrix_tophits), 0,
max(heatmap_matrix_tophits)),
c("blue", "white", "red"))
}
current_heatmap <- Heatmap(as.matrix(heatmap_matrix_tophits),
cluster_rows = TRUE,
cluster_columns = TRUE,
show_row_dend = TRUE,
show_column_dend = TRUE,
col=heatmap_col,
show_column_names = TRUE,
show_row_names = FALSE,
show_heatmap_legend = TRUE,
)print out the heat map
current_heatmap
Now we want to see how the heatmap changes after we add the group condition
top_hits <- rownames(output_hits_pat)[output_hits_pat$P.Value < 0.05]
heatmap_matrix_tophits <- scale(heatmap_matrix[which(rownames(heatmap_matrix) %in% top_hits),])
heatmap_matrix_tophits<- heatmap_matrix_tophits[,
c(grep(colnames(heatmap_matrix_tophits),
pattern = "H"),
grep(colnames(heatmap_matrix_tophits),
pattern = "D"))]
if(min(heatmap_matrix_tophits) == 0){
heatmap_col = colorRamp2(c( 0, max(heatmap_matrix_tophits)),
c( "white", "red"))
} else {
heatmap_col = colorRamp2(c(min(heatmap_matrix_tophits), 0,
max(heatmap_matrix_tophits)),
c("blue", "white", "red"))
}
current_heatmap <- Heatmap(as.matrix(heatmap_matrix_tophits),
cluster_rows = TRUE,
cluster_columns = FALSE,
show_row_dend = TRUE,
show_column_dend = FALSE,
col=heatmap_col,
show_column_names = TRUE,
show_row_names = FALSE,
show_heatmap_legend = TRUE,
)print out the heat map
current_heatmap
The data shows more clustering after we group the columns by disease, now we will redraw the heatmap with group the columns by disease, and p-value < 0.01.
top_hits <- rownames(output_hits_pat)[output_hits_pat$P.Value < 0.01]
heatmap_matrix_tophits <- scale(heatmap_matrix[which(rownames(heatmap_matrix) %in% top_hits),])
heatmap_matrix_tophits<- heatmap_matrix_tophits[,
c(grep(colnames(heatmap_matrix_tophits),
pattern = "H"),
grep(colnames(heatmap_matrix_tophits),
pattern = "D"))]
if(min(heatmap_matrix_tophits) == 0){
heatmap_col = colorRamp2(c( 0, max(heatmap_matrix_tophits)),
c( "white", "red"))
} else {
heatmap_col = colorRamp2(c(min(heatmap_matrix_tophits), 0,
max(heatmap_matrix_tophits)),
c("blue", "white", "red"))
}
current_heatmap <- Heatmap(as.matrix(heatmap_matrix_tophits),
cluster_rows = TRUE,
cluster_columns = FALSE,
show_row_dend = TRUE,
show_column_dend = FALSE,
col=heatmap_col,
show_column_names = TRUE,
show_row_names = FALSE,
show_heatmap_legend = TRUE,
)print out the heat map
current_heatmap
The colors more intense than before because we only use the genes with a p-value less than 0.01. The clusters can be clearly seen.
Question 4:
Yes, my conditions cluster together because the clusters on the heatmap can be clearly seen. We can see a clear difference between healthy and disease group after we add the group condition.
Thresholded over-representation analysis
Threshold list
Create thresholded lists using edgeR results
qlf_output_hits_withgn <- merge(filtered_data[,1],qlf_output_hits, by.x=1, by.y = 0)
qlf_output_hits_withgn[,"rank"] <- -log(qlf_output_hits_withgn$PValue,base =10) *
sign(qlf_output_hits_withgn$logFC)
qlf_output_hits_withgn <- qlf_output_hits_withgn[order(qlf_output_hits_withgn$rank),]
upregulated_genes <- qlf_output_hits_withgn$x[which(qlf_output_hits_withgn$PValue < 0.05 &
qlf_output_hits_withgn$logFC > 0)]
downregulated_genes <- qlf_output_hits_withgn$x[which(qlf_output_hits_withgn$PValue < 0.05 &
qlf_output_hits_withgn$logFC < 0)]
write.table(x=upregulated_genes,
file=file.path("data","mets_upregulated_genes.txt"),sep = "\t",
row.names = FALSE,col.names = FALSE,quote = FALSE)
write.table(x=downregulated_genes,
file=file.path("data","mets_downregulated_genes.txt"),sep = "\t",
row.names = FALSE,col.names = FALSE,quote = FALSE)
write.table(x=data.frame(genename= qlf_output_hits_withgn$x,F_stat= qlf_output_hits_withgn$rank),
file=file.path("data","mets_ranked_genelist.txt"),sep = "\t",
row.names = FALSE,col.names = FALSE,quote = FALSE)Which method did you choose and why?
I choose to use g:Profiler because it is easy to use and can give lots of information about the relative disease, it also has a lot of data sources for use. The Significance threshold was set to BH method.
What annotation data did you use and why? What version of the annotation are you using?
- GO biological process: 2020-12-08
- KEGG: KEGG 2020-12-14
- Reactome: ensembl classes: 2020-12-15
- WikiPathways: 2020-12-10
I chose to use those four dataset because it was used in homework assignment and we are interested in biological pathway and biological process
How many genesets were returned with what thresholds?
With threshold less than 0.05, there are 86 gene sets are from up regulated genes, 552 gene sets are from down regulated genes, and 543 gene sets are from differential expression data.
Run the analysis using the up-regulated set of genes, and the down-regulated set of genes separately. How do these results compare to using the whole list (i.e all differentially expressed genes together vs. the up-regulated and down regulated differentially expressed genes separately)?
The top result for up-regulated set of genes are establishment of protein localization to endoplasmic reticulum in GO:BP, Ribosome in KEGG, Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) in REAC and Cytoplasmic Ribosomal Proteins for WP. The top result for down regulated set of genes are neutrophil activation involved in immune response in GO:BP, Chemokine signaling pathway in KEGG, Neutrophil degranulation in REAC and Microglia Pathogen Phagocytosis Pathway for WP. There is not much changes after adding all deferentially expressed genes together since most of the genes are down regulated and only 5 genes are up regulated. so the top result for all deferentially expressed genes and own regulated deferentially expressed genes are same, the results are shown as below.
Interpretation
Do the over-representation results support conclusions or mechanism discussed in the original paper?
The over-representation result mechanism discussed the original paper, in the paper, up-regulated genes are at least 440, and down-regulated genes are at least 462. But in my analysis, only 5 genes are up-regulated, and 131 genes are down-regulated. This might because in the original experiment, they split the data into four groups with MetS obese, MetS lean, healthy lean, and healthy obese, and when they do comparison, they will fix one condition and compare the other condition. In my case, obese objects and lean objects are mixed together and looking for the comparison between MetS and healthy.
Can you find evidence, i.e. publications, to support some of the results that you see. How does this evidence support your results.
- Result 1: The top result for down-regulated genes are related to immune response, this could associated with MetS and obesity
support: In Andersen et al.(2016), the article stated that MetS and obesity could cause inflammation and trigger immunu responds, this evidence directly supports the finding.
- Result 2: The top result for up-regulated genes are associated with ribosomal proteins, those genes might related to obese.
support: in the original document, the RPS24 and RPL34 are both up-regulated and are associated with EIF2 Signaling. The comparison is between healthy obese and haelthy lean object, this means those genes may related to the cause of obese.
Reference
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